In light of the huge unmet need for novel antiviral strategies, an efficient solution is to identify common host factors hijacked by multiple viruses and repurpose approved drugs targeting these factors as broad-spectrum antivirals. The Khatri lab developed and validated a novel bioinformatics approach that is ideal for efficient identification of such candidate targets and antiviral drugs. By integrating meta-analysis of gene expression data and drug expression profiles we identified robust signatures of graft rejection and discovered drugs which improve graft survival. The Einav lab demonstrated feasibility of another repurposing approach. We discovered an Achilles' heel of infectious production of multiple viruses: a requirement for AP2-associated protein kinase 1 (AAK1) and cyclin G-associated kinase (GAK), host kinases that regulate clathrin adaptor proteins pathways. We then discovered inhibitors of these targets, including approved anticancer drugs; sunifinib and eriotinib, and demonstrated potent activity against hepatitis C virus, dengue virus, and HIV. We hypothesize that: 1) meta-analysis of gene expression profiles from multiple viral infections can identify common host factors critical for infection, and approved drugs targeting these factors can be used as broad-spectrum antivirals. 2) AAK1 and GAK inhibitors represent a practical novel, host centered class of broad-spectrum antivirals with utility against viruses for which no treatment is available and resistant viruses. In Aims 1 and 2 we will integrate gene expression data sets and perform meta-analysis to identify differentially expressed host factors across multiple viral infections. Existing drugs that may target these host factors will be predicted and their antiviral activity against multiple priority agents determined. Lead compounds will be further developed. Aims 3 and 4 focus on preclinical development of approved drugs with anti-AAKI and GAK activity, already displaying broad-spectrum antiviral activity, as a clinical stage product ready for testing under an IND. We will obtain data sets supporting indications for sunitinib and eriotinib against infections with multiple priority pathogens and use best available authentic models to assess the in vivo feasibility and biological rationale of this approach. These aims will integrate molecular virology, genomic, biochemical and pharmacological approaches. This project will contribute towards attainment of the multi-project center objectives by establishing a wide-spectrum repurposing pipeline against multiple NIAID Category A, B and C agents, from identifying new products at reduced time and cost to de-risking the clinical development of 2 approved drugs already showing promise.